LOCUS       sequence-243655-        4438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making C-terminal TurboID-mVenus fusion 
            with nuclear and non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4438)
  AUTHORS   Mair A, Xu SL, Branon TC, Ting AY, Bergmann DC
  TITLE     Proximity labeling of protein complexes and cell type-specific 
            organellar proteomes in Arabidopsis enabled by TurboID.
  JOURNAL   Elife. 2019 Sep 19;8. pii: 47864. doi: 10.7554/eLife.47864.
  PUBMED    31535972
REFERENCE   2  (bases 1 to 4438)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2019-09-19"; volume: "8. pii: 47864. doi"; pages: " 
            10.7554/eLife.47864"
COMMENT     Sequence Label: pDONR_P2R-P3_R2-Turbo-mVenus-STOP-L3
FEATURES             Location/Qualifiers
     source          1..4438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             786..827
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             834..1790
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             join(1827..1829,1830..1832,1833..2546)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2133..2152)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2480..2501
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2547..2642)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2679..2698)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2680..2698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2703..2719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2703..2719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2821..2840)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2832..3641
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2899..2918)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3788..4376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4277..4296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggctg ggcgcgccgg tggaggcggt tcaggaggtg
      781 gcatgggcaa gcccatcccc aaccccctgc tgggcctgga cagcaccgct agcaaagaca
      841 atactgtgcc tctgaagctg atcgctctcc tggctaatgg cgagttccat agtggcgaac
      901 agctgggaga aaccctgggc atgtccaggg ccgctatcaa caagcacatt cagactctgc
      961 gcgactgggg cgtggacgtg ttcaccgtgc ccggaaaggg ctactctctg cccgagccta
     1021 tcccgctgct gaacgctaaa cagattctgg gacagctgga cggcgggagc gtggcagtcc
     1081 tgcctgtggt cgactccacc aatcagtacc tgctggatcg aatcggcgag ctgaagagtg
     1141 gggatgcttg cattgcagaa tatcagcagg cagggagagg aagcagaggg aggaaatggt
     1201 tctctccttt tggagctaac ctgtacctga gtatgttttg gcgcctgaag cggggaccag
     1261 cagcaatcgg cctgggcccg gtcatcggaa ttgtcatggc agaagcgctg cgaaagctgg
     1321 gagcagacaa ggtgcgagtc aaatggccca atgacctgta tctgcaggat agaaagctgg
     1381 caggcatcct ggtggagctg gccggaataa caggcgatgc tgcacagatc gtcattggcg
     1441 ccgggattaa cgtggctatg aggcgcgtgg aggaaagcgt ggtcaatcag ggctggatca
     1501 cactgcagga agcagggatt aacctggaca ggaatactct ggccgctacg ctgatccgag
     1561 agctgcgggc agccctggaa ctgttcgagc aggaaggcct ggctccatat ctgccacggt
     1621 gggagaagct ggataacttc atcaatagac ccgtgaagct gatcattggg gacaaagaga
     1681 ttttcgggat tagccggggg attgataaac agggagccct gctgctggaa caggacggag
     1741 ttatcaaacc ctggatgggc ggagaaatca gtctgcggtc tgccgaaaag ggaggcggtg
     1801 gaagcggcgg aggtgggcgc gcctccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1861 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1921 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
     1981 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct
     2041 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2101 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2161 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2221 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2281 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     2341 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2401 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc
     2461 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2521 tcggcatgga cgagctgtac aagtgacaac tttattatac aaagttggca ttataaaaaa
     2581 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2641 ttggagctcc atggtagcgt taacgcggcc gcgatatccc ctatagtgag tcgtattaca
     2701 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     2761 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     2821 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     2881 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2941 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     3001 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     3061 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     3121 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     3181 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     3241 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     3301 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     3361 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     3421 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     3481 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     3541 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     3601 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     3661 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     3721 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3901 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4141 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     4201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
